Introduction
The Kit provides fast purification of high-quality RNA from paxigene Blood RNA Tube using silica-membrane spin columns. RNA purified using the HiPure System is ready for applications such as RT-PCR, Northern blotting, poly A+ RNA (mRNA) purification, nuclease protection, and in vitro translation.
Details
Specifications
| Features | Specifications |
| Main Functions | Isolation total RNA from Paxgene RNA Tubes |
| Applications | RT-PCR, qPCR, Northern hybridization, NGS, nucleic acid protection, in vitro translation |
| Purification method | Mini spin column |
| Purification technology | Silica technology, DNA filtration technology |
| Process method | Manual (centrifugation or vacuum) |
| Sample type | Blood in the preservation tube |
| Sample amount | 2.5ml |
| Elution volume | ≥30μl |
| Liquid carrying volume per column | 800µl |
| Binding yield of column | 100μg |
Principle
The Kit is for the purification of total RNA from 2.5 ml human whole blood collected in a PAXgene Blood RNA Tube. Purification begins with a centrifugation step to pellet nucleic acids in the PAXgene Blood RNA Tube. The pellet is washed and resuspended, and incubated in optimized buffers together with proteinase K. DNA wash The lysate is passed through a DNA Mini column. Ethanolis added to adjust binding conditions, and the lysate is applied to a column.RNA is selectively bound to the silica membrane as contaminants pass through. Remaining contaminants are removed in several efficient wash steps. Between the first and second wash steps, the membrane is treated with DNase I to remove trace amounts of bound DNA. After the wash steps, RNA is eluted in elution buffer and heat-denatured.
Advantages
- High quality – high purity total RNA can be directly used in various sensitive downstream applications
- Fast – isolation of several samples can be completed in 40 minutes by using column purification method
- Safety – no phenol chloroform extraction required
- Sensitive – direct lysis of blood, plasma and other samples without separation of leukocytes
Kit Contents
| Contents | R416802 | R416803 |
| Purification Times | 10 Preps | 50 Preps |
| HiPure RNA Mini Columns I | 10 | 50 |
| gDNA Filter Mini Columns | 10 | 50 |
| 2ml Collection Tubes | 30 | 150 |
| RNase Free Water | 60 ml | 250 ml |
| Buffer MBR1 | 10 ml | 30 ml |
| Buffer MBR2 | 5 ml | 15 ml |
| Buffer RW1 | 15 ml | 60 ml |
| Buffer RW2* | 6 ml | 20 ml |
| Proteinase K | 12 mg | 50 mg |
| Protease Dissolve Buffer | 1.8 ml | 5 ml |
| DNase I | 120 µl | 600 µl |
| DNase Buffer | 6 ml | 30 ml |
Storage and Stability
Proteinase K should be stored at 2–8°C upon arrival. DNase I should be stored at -20°C. However, short-term storage (DNase I up to 1 weeks, Proteinase K up to 8 weeks) at room temperature (15–25°C) does not affect their performance. The remaining kit components can be stored at room temperature (15–25°C) and are stable for at least 18 months under these conditions.
Experiment Data
